Evaluation of IRES-mediated, cell-type-specific cytotoxicity of poliovirus using a colorimetric cell proliferation assay.
PVS-RIPO is a recombinant oncolytic poliovirus designed for clinical application to target CD155 expressing malignant gliomas and other malignant diseases. PVS-RIPO does not replicate in healthy neurons and is therefore non-pathogenic in rodent and non-human primate models of poliomyelitis. A tetrazolium salt dye-based cellular assay was developed and qualified to define the cytotoxicity of virus preparations on susceptible cells and to explore the target cell specificity of PVS-RIPO. In this assay, PVS-RIPO inhibited proliferation of U87-MG astrocytoma cells in a dose-dependent manner. However, HEK293 cells were much less susceptible to cell killing by PVS-RIPO. In contrast, the Sabin type 1 live attenuated poliovirus vaccine strain (PV(1)S) was cytotoxic to both HEK293 and U87-MG cells. The correlation between expression of CD155 and cytotoxicity was also explored using six different cell lines. There was little or no expression of CD155 and PVS-RIPO-induced cytotoxicity in Jurkat and Daudi cells. HEK293 was the only cell line tested that showed CD155 expression and resistance to PVS-RIPO cytotoxicity. The results indicate that differential cytotoxicity measured by the colorimetric assay can be used to evaluate the cytotoxicity and cell-type specificity of recombinant strains of poliovirus and to demonstrate lot to lot consistency during the manufacture of viruses intended for clinical use.
Yang
Xiaoyi
X
Bioanalytical Development Laboratory, Biopharmaceutical Development Program, SAIC-Frederick, Inc., NCI Frederick, USA.
Chen
Eying
E
Jiang
Hengguang
H
Muszynski
Karen
K
Harris
Raymond D
RD
Giardina
Steven L
SL
Gromeier
Matthias
M
Mitra
Gautam
G
Soman
Gopalan
G
D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003124:Colorimetry / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005909:Glioblastoma; D006801:Humans; D009457:Neuroglia / Q000821:virology*; D017955:Poliovirus / Q000472:pathogenicity*; D011055:Poliovirus Vaccine, Oral; D011991:Receptors, Virus / Q000378:metabolism
D011055:Poliovirus Vaccine, Oral; D011991:Receptors, Virus; C066100:poliovirus receptor